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Updated: Aug 11, 2025
Inducing Acute Lung Injury in Mice by Direct Intratracheal Lipopolysaccharide Instillation
Published on: July 6, 2019
Morita-Baylis-Hillman adduct 2-(3-hydroxy-2-oxoindolin-3-yl)acrylonitrile (ISACN) modulates the inflammatory process
Juliane Santos de França da Silva1, Deyse Cristina Madruga Carvalho2, Luiz Henrique Agra Cavalcante-Silva2
1Centro de Ciências da Saúde, Universidade Federal da Paraíba, João Pessoa, Brasil.
Background:
Despite its homeostatic role, inflammation is involved in several pathologies, such as acute lung injury. Morita-Ballys-Hilman adducts (MBHA) are a group of synthetic molecules and present a wide range of biological activities, including anti-inflammatory action. Thus, this study aimed to assess whether ISACN, an MBHA, modulates inflammation during acute lung injury induced by lipopolysaccharide (LPS).
Methods:
BALB/c mice were intraperitoneally treated with 24 mg/kg ISACN and challenged with LPS (2.5 mg/kg). On bronchoalveolar lavage fluid (BALF), we assessed the total and differential leukocyte count and measurement of protein leakage, cytokines (IL-1β, IL-6, and TNF-α), and chemokine (CXCL-1). Additionally, lung histopathology was also performed (H&E staining). In vitro studies were conducted with peritoneal macrophages to assess the possible mechanism of action. They were cultured in the presence of ISACN (5 and 10 µM) and stimulated by LPS (1 µg/mL).
Results:
ISACN reduced neutrophil migration, protein leakage, and inflammatory cytokines (IL-1β, IL-6, and TNF-α) without interfering with the production of CXCL1. In addition, ISACN caused a decrease in LPS-induced lung injury as evident from histopathological changes. In peritoneal macrophages, ISACN diminishes the nitric oxide and cytokine levels (IL-1β, IL-6, and TNF-α). The treatment with ISACN (10 μM) also reduced LPS-induced TLR4, CD69, iNOS overexpression, and the LPS-induced ERK, JNK, and p38 phosphorylation.
Conclusion:
Thus, this work showed for the first time the immunomodulatory action of MBHA in LPS-induced acute lung injury and provided new evidence for the mechanisms related to the anti-inflammatory effect of ISACN.
Insights
This study demonstrates that ISACN, a Morita-Baylis-Hillman adduct (MBHA), effectively reduces inflammation and lung injury caused by lipopolysaccharide (LPS). ISACN modulates immune responses, offering potential therapeutic benefits for acute lung injury.
Area of Science:
- Immunology
- Pharmacology
- Toxicology
Background:
- Inflammation plays a key role in pathologies like acute lung injury.
- Morita-Baylis-Hillman adducts (MBHA) are synthetic molecules with known anti-inflammatory properties.
- ISACN is an MBHA investigated for its potential to modulate inflammation.
Purpose of the Study:
- To evaluate the anti-inflammatory effects of ISACN on lipopolysaccharide (LPS)-induced acute lung injury.
- To investigate the underlying mechanisms of ISACN's action in modulating inflammation.
Main Methods:
- BALB/c mice were treated with ISACN and challenged with LPS.
- Bronchoalveolar lavage fluid (BALF) analysis included leukocyte counts, protein leakage, and cytokine/chemokine levels.
- Lung histopathology and in vitro studies on peritoneal macrophages were performed.
Main Results:
- ISACN significantly reduced neutrophil migration, protein leakage, and inflammatory cytokines (IL-1β, IL-6, TNF-α) in BALF.
- Histopathological examination showed decreased LPS-induced lung injury.
- In vitro, ISACN reduced nitric oxide, cytokine production, TLR4, CD69, iNOS overexpression, and MAPK phosphorylation in macrophages.
Conclusions:
- ISACN exhibits immunomodulatory action in LPS-induced acute lung injury.
- This study provides novel insights into the anti-inflammatory mechanisms of ISACN, highlighting its therapeutic potential.
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